Dcm methylation is detrimental to plasmid transformation in Clostridium thermocellum.

Biotechnology for Biofuels
Adam M GussLee R Lynd

Abstract

Industrial production of biofuels and other products by cellulolytic microorganisms is of interest but hindered by the nascent state of genetic tools. Although a genetic system for Clostridium thermocellum DSM1313 has recently been developed, available methods achieve relatively low efficiency and similar plasmids can transform C. thermocellum at dramatically different efficiencies. We report an increase in transformation efficiency of C. thermocellum for a variety of plasmids by using DNA that has been methylated by Escherichia coli Dam but not Dcm methylases. When isolated from a dam+dcm+E. coli strain, pAMG206 transforms C. thermocellum 100-fold better than the similar plasmid pAMG205, which contains an additional Dcm methylation site in the pyrF gene. Upon removal of Dcm methylation, transformation with pAMG206 showed a four- to seven-fold increase in efficiency; however, transformation efficiency of pAMG205 increased 500-fold. Removal of the Dcm methylation site from the pAMG205 pyrF gene via silent mutation resulted in increased transformation efficiencies equivalent to that of pAMG206. Upon proper methylation, transformation efficiency of plasmids bearing the pMK3 and pB6A origins of replication increased ca. three orde...Continue Reading

References

Jan 1, 1991·Methods in Enzymology·T Kieser, D A Hopwood
Feb 1, 1991·Journal of Bacteriology·A Macaluso, A M Mettus
Mar 1, 1993·Research in Microbiology·A A VertèsH Yukawa
Mar 1, 1996·Applied Microbiology and Biotechnology·T R KlapatchL R Lynd
Sep 5, 2002·Microbiology and Molecular Biology Reviews : MMBR·Lee R LyndIsak S Pretorius
Mar 26, 2003·Nucleic Acids Research·Richard J RobertsShuang-yong Xu
Feb 10, 2004·Applied and Environmental Microbiology·Michael V TyurinLee R Lynd
Jul 6, 2006·Applied and Environmental Microbiology·Robert M Q ShanksGeorge A O'Toole
Feb 9, 2008·Nature Biotechnology·Lee R LyndCharles E Wyman
Oct 23, 2009·Nucleic Acids Research·Richard J RobertsDana Macelis
Aug 10, 2010·Applied and Environmental Microbiology·Shital A TripathiNicky C Caiazza
Sep 15, 2010·Proceedings of the National Academy of Sciences of the United States of America·Daniel G OlsonLee R Lynd
Apr 5, 2011·Journal of Bacteriology·Lawrence FeinbergNicky Caiazza

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Citations

Jul 15, 2015·Metabolic Engineering·Paul P LinJames C Liao
May 23, 2015·Applied Microbiology and Biotechnology·Elizabeth B MearlsLee R Lynd
Jul 15, 2015·Journal of Industrial Microbiology & Biotechnology·Thomas RydzakAdam M Guss
Oct 15, 2013·FEMS Microbiology Reviews·Sara E Blumer-SchuetteRobert M Kelly
Jun 4, 2016·Biotechnology for Biofuels·Liang TianLee R Lynd
Apr 11, 2018·Biotechnology for Biofuels·Jason M WhithamSteven D Brown
Feb 2, 2018·Journal of Basic Microbiology·Yayun WangKazuo Sakka
May 12, 2018·Biotechnology and Bioengineering·Emily FreedCarrie A Eckert
Dec 23, 2016·Applied and Environmental Microbiology·Charlotte M WilsonSteven D Brown
Oct 6, 2017·Microbial Cell Factories·Liang TianLee R Lynd
Jul 25, 2018·Scientific Reports·Lizett Ortiz de OraIván Muñoz-Gutiérrez
Jan 6, 2017·Biotechnology for Biofuels·Ranjita BiswasAdam M Guss
Jan 27, 2021·Biotechnology for Biofuels·Lauren A Riley, Adam M Guss

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Methods Mentioned

BETA
genetic modification
PCR

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